Cheng Hongxia, Cui Xinhua, Guo Ying, Gu Lintao, Wang Yaning, Wang Qirong, Liang Hui
Department of Pathology, Provincial Hospital Affiliated to Shandong University 324, Jing 5 Rd, Jinan 250021, Shandong, China.
Department of Otolaryngology, Shandong Provincial Qianfoshan Hospital 16766, Jingshi Road, Jinan 250014, Shandong, China.
Am J Transl Res. 2016 Nov 15;8(11):5052-5058. eCollection 2016.
Human hypopharyngeal carcinoma is one of the most common malignant tumors. CD44 could serve as a molecular marker to screen for cancer stem cells (CSCs) in hypopharyngeal cancer. The aim of this study was to identify the role of HOX transcript antisense RNA (HOTAIR) on cell proliferation and invasion in CD44+ FADU cells (human hypopharyngeal carcinoma cells). We also explored the underlying mechanism contributing to HOTAIR's observed effects. CD44+ FADU cells were sorted and purified by flow cytometry and infected with lentivirus stably expressing HOTAIR shRNA. Cell proliferation and invasion analyses were carried out with cell counting kit-8 (CCK-8) and Transwell assays. The expressions of downstream effectors of HOTAIR, including E-cadherin, β-catenin, and vimentin were measured by quantitative real-time polymerase chain reaction (qRT-PCR) and western blotting. Knockdown of HOTAIR markedly inhibited the proliferation and invasion of CD44+ FADU cells . HOTAIR depletion also increased the expressions of tumor suppressors E-cadherin and β-catenin and decreased the expression of oncogenic vimentin at both mRNA and protein levels. Collectively, our results show that HOTAIR can suppress CD44+ FADU cells proliferation and invasion by regulating the expressions of E-cadherin, β-catenin, and vimentin.
人下咽癌是最常见的恶性肿瘤之一。CD44可作为筛选下咽癌中癌症干细胞(CSCs)的分子标志物。本研究的目的是确定HOX转录本反义RNA(HOTAIR)在CD44+FADU细胞(人下咽癌细胞)的细胞增殖和侵袭中的作用。我们还探讨了导致HOTAIR观察到的效应的潜在机制。通过流式细胞术对CD44+FADU细胞进行分选和纯化,并用稳定表达HOTAIR shRNA的慢病毒进行感染。使用细胞计数试剂盒-8(CCK-8)和Transwell实验进行细胞增殖和侵袭分析。通过定量实时聚合酶链反应(qRT-PCR)和蛋白质印迹法检测HOTAIR下游效应分子E-钙黏蛋白、β-连环蛋白和波形蛋白的表达。敲低HOTAIR可显著抑制CD44+FADU细胞的增殖和侵袭。HOTAIR的缺失还在mRNA和蛋白质水平上增加了肿瘤抑制因子E-钙黏蛋白和β-连环蛋白的表达,并降低了致癌性波形蛋白的表达。总的来说,我们的结果表明,HOTAIR可通过调节E-钙黏蛋白、β-连环蛋白和波形蛋白的表达来抑制CD44+FADU细胞的增殖和侵袭。